Investigation of hemostatic effect of Spleeninvigorating,Qi-replenishing and Bloodarresting Formula on simvastatin-induced zebrafish hemorrhage model

Blood-arresting Formula on simvastatin-induced zebrafish hemorrhage model compared with that of the Heat-clearing and Blood-cooling Formula.Methods:AB strain wild type zebrafish were treated with 0.5 mM simvastatin for 24 hours to establish the zebrafish hemorrhage model.Under strictly blind experimental conditions,the zebrafish were then treated with experimental agents in different concentration within the maximum non-lethal dose.The intervention effect of the Spleen-invigorating,Qi-replenishing and Blood-arresting Formula was comprehensively assessed by examining the main observational parameters such as the bleeding reduction rate and hemostatic rate while referring to the additional parameters such as blood flow,improvement rate of blood flow,velocity of movement,improvement rate of motion under spleen-qi deficiency.Results:Hemostatic effect:The bleeding rate and hemostatic rate in the zebrafish were respectively,30%,15%,45%,40%and 60%,80%,40%,47%when equal concentration of 500 mg/mL,1000 mg/mL were used.This showed that the experimental herbs B1 was superior to B2 in decreasing the bleeding rate and improving hemostatic effect in the zebrafish.Improvement of blood flow:With equal concentration,the experimental herb B1 was superior to B2 in increasing the blood flow and improvement rate of blood flow in hemorrhagic zebrafish.Promotion of the improvement of motion:With equal concentration of 500 mg/mL and 1000 mg/mL,the experimental herb B1 was superior to B2 in promoting the velocity of motion in the zebrafish and promoting the improvement rate of motion.Conclusion:The manifestations of simvastatin-induced zebrafish hemorrhage are basically similar to that of failure of the spleen to control blood in human beings.The Spleeninvigorating,Qi-replenishing and Blood-arresting Formula has good hemostatic effect on simvastatin-induced hemorrhage in the zebrafish.It also boasts the functions of improving blood flow and velocity of motion in the hemorrhagic zebrafish.Such a conclusion provides the experimental basis for the treatment of the syndrome of failure of the spleen to control blood by this formula.

Effect of spleen-invigorating, Qi-replenishing and blood-arresting formula on zebrafish models with simvastatin-induced hemorrhage caused by spleen failing to control blood, in terms of theory of Traditional Chinese Medicine

OBJECTIVE: To investigate the hemostasis effect of spleen-invigorating, Qi-replenishing and blood-arresting formula, on a zebrafish models with simvastatin-induced hemorrhage, and with symptom pattern caused by spleen failing to control blood, in terms of theory of Traditional Chinese Medicine(TCM).METHODS: In the first experiment, 60 AB strain wild type zebrafishes were randomly assigned into two groups: normal group and model group. The model group was treated with 50 μM simvastatinfor 24 h. The second experiment: The melanin allele mutated Albino strain zebrafishes were divided into normal, model, A group and B group. The observational parameters were as follows: blood flow, velocity of movement, hemorrhage ratio and improvement ratio of hemorrhage.RESULTS: Hemorrhage ratio: in the first experiment, brain hemorrhage ratio was 75%. In the second experiment, heart hemorrhage ratio was 65%.Blood flow: compared with the normal group,there was a significantly decrease in the model group(P < 0.001). Velocity of movement: in the first experimental, compared with the normal group,there was a significantly decrease in the model group(P < 0.001). Improvement ratio of hemorrhage: agents A had little effect in heart hemorrhage of the zebrafish; agents B could reduce heart hemorrhage ratio of the zebrafish, and increase the improvement ratio of hemorrhage.CONCLUSION: The manifestation of zebrafish model with simvastatin-induced hemorrhage is basically similar to that of the clinical symptom pattern caused by spleen's failure to control blood. The Spleen-invigorating, Qi-replenishing and Blood-arresting Formula can reduce the heart hemorrhage ratio of zebrafish induced by simvastatin, and increase the Improvement ratio of hemorrhage.